糖基化终产物对THP-1巨噬细胞酰基辅酶A:胆固醇酰基转移酶-1表达的影响

Effects of advanced glycation end-products on expression of acyl coenzyme A:cholesterol acyltransferase-1 in THP-1 macrophages

目的观察糖基化终产物(advanced glycosylation end-products,AGEs)对THP-1巨噬细胞酰基辅酶A:胆固醇酰基转移酶-1(acyl coenzyme A:cholesterol acyltransferase-1,ACAT-1)表达的影响。方法将糖基化-牛血清白蛋白(AGE-modified bovine serum albumin,AGE-BSA)与佛波酯(PMA)诱导分化72h的THP-1巨噬细胞共同孵育,运用RT-PCR和Western blot分别检测巨噬细胞ACAT-1 mRNA和蛋白的表达水平。结果PMA诱导72h后,THP-1细胞停止增殖,由单核细胞分化成为巨噬细胞。用50、100、200和400mg/L AGE-BSA处理巨噬细胞后,细胞ACAT-1 mRNA相对表达量逐渐增加;ACAT-1蛋白表达的平均积分光密度值也逐渐增加,二者较对照牛血清白蛋白组明显增加(P<0.05)。用200mg/LAGE-BSA作用巨噬细胞0、12、24、36h和48h后,细胞ACAT-1 mRNA相对表达量逐渐升高;ACAT-1蛋白表达的平均积分光密度值也逐渐升高,较0h明显增加(P<0.05)。结论AGEs可上调THP-1巨噬细胞ACAT-1 mRNA和蛋白的表达,呈浓度和时间依赖性,这可能与糖尿病动脉粥样硬化有关。

Objective To investigate the effects of advanced glycosylation end-products （AGEs） on the expression of acyl coenzyme A： cholesterol acyhransferase-1 （ ACAT-1 ） in cuhured THP-1 macrophages. Methods THP-1 cells were differentiated into macrophages after cultured in RPMI1640 media containing 0.1 μmoL/L PMA for 72 h. THP-1 macrophages were then exposed to AGE-modified bovine serum albumin （AGE- BSA, at concentration of 50, 100, 200 mg/L） for 24 h or to 200 mg/L AGE-BSA for 0, 12, 24, 36 h. Expression of ACAT-1 mRNA and protein in THP-1 macrophages was measured by RT-PCR and Western blot, respectively. Results After induced by 0.1μmol/L PMA, THP-1 cells stopped proliferation and differentiated into macrophages. Treatment of THP-1 macrophages with AGE-BSA resulted in an increase in the mRNA and protein levels for ACAT-1 in a doseand time-dependent manner. Conclusion AGEs upregulate the expression levels of mRNA and protein for ACAT-1 in cultured THP-1 macrophages, which might be partly involved in the atherogenesis in diabetic patients.